HIV VARIATION AND CROSS CLADE CTL RECOGNITION

描述：（改编自申请人的摘要）本项目是响应 计划公告# PA-98-011“HIV变异对免疫系统的影响” 识别.“这项提案的目的是调查跨进化枝艾滋病毒 特异性CTL应答。刺激广泛的交叉进化枝反应性HIV 特异性CTL应答被认为是成功的HIV疫苗的关键。的 高度序列变异是HIV感染的显著特征， 这是疫苗开发的一大障碍。然而，很少有信息 已知HIV特异性CTL应答在不同的国际人群中 HLA类型，感染HIV进化枝变异体。通过我们的合作伙伴，我们 获得来自非洲、中国、南非、非洲和非洲艾滋病毒感染者的PBMC样本 美国和泰国感染HIV-1和HIV-2。在第一个具体目标中， 我们将确定是否存在交叉反应性CTL表位， 感染不同分支的HIV-1的人群。我们将测量交叉 用敏感的ELISPOT试验检测反应性CTL应答， 表达HIV-1基因产物的进化枝特异性重组痘苗病毒， env、gag、p01和nef以及进化枝特异性肽。原液培养物CTL应答 将通过多克隆和抗原特异性再刺激生成并进行检测 对自体或匹配的B淋巴母细胞样细胞靶点具有进化枝特异性 抗原我们将把CTL频率与所有受试者的病毒RNA血浆载量相关联， 进化枝，并评估氨基酸变化是否在最佳定义的最小 表位可以使HIV CTL表位更具交叉反应性。在第二个具体 目的：探讨HlV~2感染是否能刺激CTL产生交叉反应， 与HIV-1反应或反之亦然，以及一些人是否受到保护， HIV-1的一个进化枝对抗另一个进化枝的重叠感染。

DESCRIPTION: (Adapted from Applicant's Abstract) This project is responsive to the Program Announcement # PA-98-011 "Impact of HIV Variation on Immunological Recognition." The aim of this proposal is to investigate cross clade HIV specific CTL responses. Stimulation of a broad cross clade reactive HIV specific CTL response is considered crucial for a successful HIV vaccine. The high degree of sequence variation is a prominent feature of HIV infection, and a major impediment to vaccine development. However, very little information is known about HIV specific CTL responses in international populations of diverse HLA types, infected with HIV clade variants. Through our collaborators, we have access to PBMC samples from HIV infected individuals in Africa, China, South America and Thailand infected with HIV-1 and HIV-2. In the first specific aim, we will determine whether cross-reactive CTL epitopes exist in diverse populations infected with different clades of HIV-1. We will measure cross reactive CTL responses with a sensitive ELISPOT assay, using both a panel of clade specific recombinant vaccinia viruses expressing HIV-1 gene products from env, gag, p01 and nef, and clade specific peptides. Bulk culture CTL responses will be generated with polyclonal and antigen specific restimulation and tested on autologous or matched B lymphoblastoid cell targets with clade specific antigens. We will correlate CTL frequency with plasma load of viral RNA for all clades and assess whether amino acid changes in optimally defined minimal epitopes can make HIV CTL epitopes more cross reactive. In the second specific aim, we will ascertain whether infection with HlV~2 stimulates CTL which cross react with HIV-1 or vice versa, and whether some individuals are protected by one clade of HIV-1 against super-infection with another.